Visualization of necroptotic cell death through transmission electron microscopy

Naresh Golla, Linda J. Hong, Ilana Chefetz

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Transmission electron microscopy (TEM) is an all-in-one tool to visualize the complex systems of any specimen that is 1 nm in size or smaller. The current chapter provides detailed guidelines for imaging morphological changes during programmed cell necrosis using TEM as a single-step methodology. In this protocol, a novel aldehyde dehydrogenase inhibitor is used to induce cell programmed necrosis in ovarian cancer cell lines (A2780 and SKOV3). This process is followed by gradient dehydration with ethanol, chemical fixation, sampled grid preparation, and staining with 0.75% uranyl formate. Following fixation and grid preparation, cells are imaged using TEM. The resulting images reveal morphological changes consistent with necrotic morphology, including swelling of cells and organelles, appearance of vacuoles, and plasma membrane rupture followed by leakage of cellular contents. The current approach allows a single-step methodology for characterization of cell-programmed necrosis in cells based on morphology.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages135-147
Number of pages13
Volume2255
DOIs
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2255
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Publisher Copyright:
© 2021, Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

  • Aldehyde dehydrogenase inhibition
  • Cellular morphology
  • Necroptosis
  • Negative staining
  • Transmission electron microscopy (TEM)

PubMed: MeSH publication types

  • Journal Article

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